利用探针改变链自组装反应技术对反义寡核苷酸进行灵敏和特异的定量分析。

IF 2.2 4区工程技术 Q3 BIOCHEMICAL RESEARCH METHODS

BioTechniques Pub Date : 2023-07-01 DOI:10.2144/btn-2023-0005

Masako Osawa, Takurou Akiya, Funa Ogawa, Takao Suzuki, Masaki Yamagami, Tadashi Umemoto, Akira Ideno

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引用次数: 0

摘要

在药物开发过程中建立药代动力学特性时，定量生物分析是必不可少的。为了克服反义寡核苷酸(ASOs)传统分析的敏感性、特异性和过程复杂性的挑战，研究了一种利用探针改变连接自组装反应(PALSAR)技术作为信号放大器进行非酶杂交分析的新方法。PALSAR定量小鼠组织和血浆中ASOs的灵敏度为1.5 ~ 6 pg/ml，日内/日间准确度分别为86.8 ~ 119.1%和88.1 ~ 113.1%，精密度≤17.2%。此外，具有单碱基差异的代谢物3′n-1的交叉反应性也较高

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Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technology.

Quantitative bioanalysis is essential when establishing pharmacokinetic properties during the drug development process. To overcome challenges of sensitivity, specificity and process complexity associated with the conventional analysis of antisense oligonucleotides (ASOs), a new approach to nonenzymatic hybridization assays using probe alteration-linked self-assembly reaction (PALSAR) technology as a signal amplifier was evaluated. PALSAR quantification of ASOs in mouse tissue and plasma was able to achieve a high sensitivity ranging from 1.5 to 6 pg/ml, intra-/interday accuracies in the range of 86.8-119.1% and 88.1-113.1%, respectively, and precision of ≤17.2%. Furthermore, crossreactivity of 3'n-1, a metabolite with a single base difference, was <1%. Our approach provides an auspicious method for distinguishing metabolites and detecting ASOs with high sensitivity and specificity.

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来源期刊

BioTechniques 工程技术-生化研究方法

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

3.3 months

期刊介绍： BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.